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Biomedical subjects

S Kawauchi

Publications and source records attributed to S Kawauchi.

90 records · Page 5Linked to original sources
PubMed1984

Bindings of Ca2+ and substrate analogs to a cobra venom phospholipase A2 in which the alpha-amino group is modified to an alpha-keto group.

The pH dependence of the chemical reaction rate of p-bromophenacyl bromide (BPB) with His 48 of cobra (Naja naja atra) venom phospholipase A2, in which the alpha-NH2 group had been selectively modified to an alpha-keto group, was studied at 25 degrees C and ionic strength 0.1 in the absence of Ca2+. The pH-dependence curve was monophasic with a midpoint at pH 7.9, which corresponds to the pK value of His 48 of the alpha-NH2-modified enzyme, whereas the curve for the intact enzyme was biphasic, indicating participation of two ionizable groups with pK values of 7.3 and 8.55 (Teshima et al. (1982) J. Biochem. 91, 1778-1788). These two groups were thus identified as His 48 and the alpha-NH2 group, respectively. The pH dependence of the binding constant of Ca2+ to the alpha-NH2-modified enzyme was studied at 25 degrees C and ionic strength 0.1 by measuring the tryptophyl fluorescence changes. The pH-dependence curve was very similar to that for the intact enzyme (Teshima et al. (1981) J. Biochem. 89, 13-20), and it was interpreted in terms of participation of His 48 and Asp 49 (pK 5.4). The absence of participation of the alpha-NH2 group in the Ca2+ binding was thus confirmed. Bindings of monodispersed n-dodecylphosphorylcholine (n-C12PC) and micellar n-hexadecylphosphorylcholine (n-C16PC) to the alpha-NH2-modified enzyme were studied at 25 degrees C and ionic strength 0.1 by the aromatic circular dichroism (CD) and tryptophyl fluorescence methods, respectively. The binding constant of the monodispersed substrate was very similar to that for the intact enzyme (Teshima et al. (1981) J. Biochem. 89, 1163-1174). The binding constant of the micellar substrate to the modified enzyme in the presence of Ca2+ was also very similar to that for the intact enzyme-Ca2+ complex (Teshima et al. (1983) J. Biochem. 94, 223-232), and the pH-dependence curve was interpreted in terms of participation of His 48. On the other hand, the binding constant of the micellar substrate to the modified apoenzyme was much smaller than that for the intact apoenzyme. Nevertheless, the pH-dependence curve could be interpreted in terms of participation of His 48 and Asp 49. From these findings, it was concluded that the ionization state of the alpha-NH2 group of cobra venom phospholipase A2 is essentially irrelevant to the bindings of Ca2+ and also of the monodispersed and micellar substrates.

Animals↗
PubMed1982

Role of phospholipids in the inhibitory action of DDT and permethrin on the nerve ATPase of lobster, Homarus americanus.

Efforts were made to understand the nature of the site of 1,1-bis-(p-chlorophenyl)-2,2,2-trichloroethane (DDT) inhibition of nerve ATPase. THe phospholipid content of nerve preparations from the walking leg of the lobster was reduced by treating them with phospholipase A, or with a chloroform-methanol mixture at -75 degrees. By these treatments the enzymes lost approximately 70 or 95% of their phospholipids and 50-80% of their Na,K- and Ca-ATPase activities. The lost ATPase activities could be partially restored by the addition of phospholipids, either the ones extracted from the lobster nerves or those from commercial sources. ATPase inhibition by DDT and permethrin was found to be highest in preparations where the phospholipids were removed by the above treatments, next highest with the untreated original enzyme, and least with the reconstituted ATPase regardless of the source of phospholipids used for reconstitution. This tendency was more pronounced in the case of Ca-ATPase. The effects of DDT and permethrin on inhibition of reconstituted Ca-ATPase were higher when the insecticide was first added to the protein portion and the enzyme was then reconstituted with the phospholipids, than when the same amount of insecticide was first added to the phospholipids which were then used for reconstitution.

Adenosine Triphosphatases↗
PubMed1981

Properties of the mercury and selenium complex formed in rat plasma in vivo.

Properties of the mercury and selenium complex (Hg-Se Complex) produced in plasma after the simultaneous injection of mercuric chloride and sodium selenite were investigated by means of pronase digestion and isoelectric focusing. When the gel chromatographic fraction containing the HG-Se complex was digested with Pronase E and chromatographed on a Sephadex G-200 column, the resulting derived proteins contained no mercury and very low percentage of selenium. Most of mercury and selenium in the digest of the Hg-Se complex remained at the top of column, but the content of proteins (derived proteins) found at the same site of column was only trace amount. In contrast, the derived proteins obtained from proteins containing mercury or selenium in serum by the same digestion contained mercury and high percentage of selenium respectively. These results indicated that the Hg-Se complex in plasma did not contain protein as a component. When the isoelectric focusing of the serum containing the Hg-Se complex was carried out, the peak containing both mercury and selenium was present in the position of pH2 and pH4. The former peak was independent of the serum proteins, the latter peak overlapped the distribution of that. Similar behavior of mercury and selenium was observed in the isoelectric focusing of the mercuric selenide colloid dispersed in the solution containing bovine serum albumin. The properties of the Hg-Se complex in these studies supported the hypothesis that it is a mercuric selenide colloid.

Animals↗
PubMedDate not supplied

Bicarbonate stimulatory action of nizatidine, a histamine H(2)-receptor antagonist, in rat duodenums.

Nizatidine, a histamine H(2)-antagonist, is known to inhibit acetylcholinesterase (AChE) activity and is used clinically as a gastroprokinetic agent as well as the anti-ulcer agent. We examined whether or not nizatidine stimulates duodenal HCO(3)(-) secretion in rats through vagal-cholinergic mechanisms by inhibiting AChE activity. Under pentobarbital anesthesia, a proximal duodenal loop was perfused with saline, and the HCO(3)(-) secretion was measured at pH 7.0 using a pH-stat method and by adding 10 mM HCl. Nizatidine, neostigmine, carbachol, famotidine or ranitidine was administered i.v. as a single injection. Intravenous administration of nizatidine (3-30 mg/kg) dose-dependently increased the HCO(3)(-) secretion, and the effect at 10 mg/kg was equivalent to that obtained by carbachol at 0.01 mg/kg. The HCO(3)(-) stimulatory action of nizatidine was observed at the doses that inhibited the histamine-induced acid secretion and enhanced gastric motility. This effect was mimicked by neostigmine (0.03 mg/kg) and significantly attenuated by bilateral vagotomy and pretreatment with atropine but not indomethacin. The IC(50) of nizatidine for AChE of rat erythrocytes was 1.4 x 10(-6) M, about 12 times higher than that of neostigmine. Ranitidine showed the anti-AchE activity and increased duodenal HCO(3)(-) secretion, similar to nizatidine, whereas famotidine had any influence on neither AChE activity nor the HCO(3)(-) secretion. On the other hand, duodenal damage induced by acid perfusion (100 mM HCl for 4 h) in the presence of indomethacin was significantly prevented by nizatidine and neostigmine, at the doses that increased the HCO(3)(-) secretion. These results suggest that nizatidine increases HCO(3)(-) secretion in the rat duodenum, mediated by vagal-cholinergic mechanism, the action being associated with the anti-AChE activity of this agent.

Acetylcholinesterase↗
PubMedDate not supplied

Gastric acid secretion in streptozotocin-diabetic rats--different responses to various secretagogues.

We compared gastric acid secretion in response to various stimuli in normal and streptozotocin (STZ)-induced diabetic rats, in an attempt to characterize the alteration of acid secretory response in diabetic conditions. Animals were injected STZ (70 mg x kg(-1), i.p.) and used after 5 weeks of diabetes with blood glucose > 350 mg x dL(-1). Under urethane anesthesia, a rat stomach was mounted on an ex vivo chamber, perfused with saline and acid secretion was measured at pH 7.0 using a pH-stat method and by adding 100 mM NaOH. The acid secretion was stimulated by i.v. infusion of either histamine (4 mg x kg(-1) x h(-1)), pentagastrin (60 microg x kg(-1) x h(-1)) or carbachol (20 microg x kg(-1) x h(-1)) or i.v. injection of YM-14673 (0.3 mg x kg(-1)), an analog of thyrotropin-releasing hormone, or vagal electrical stimulation (2 ms, 3 Hz, 0.5 mA). In normal rats, gastric acid secretion was increased in response to either histamine, pentagastrin, carbachol, YM-14673 or electrical vagal stimulation. In STZ diabetic rats, however, changes in acid secretion varied depending on the stimuli; the acid secretory responses to histamine remained unchanged, those to YM-14673 and vagal electrical stimulation significantly decreased, but the responses to both pentagastrin and carbachol were significantly enhanced as compared to normal rats. Luminal release of histamine in response to both pentagastrin and carbachol was increased in STZ-diabetic rats as compared to normal animals. The altered acid secretory responses in STZ diabetic rats were partially reversed by daily injection of insulin with amelioration of high blood glucose levels. These results suggest that STZ-diabetic rats showed different changes in gastric acid secretory responses to various stimuli; no change in response to histamine, a decrease to both YM-14673 and vagal electrical stimulation and an increase to both pentagastrin and carbachol. The increased acid secretory response may be associated with an enhanced release of mucosal histamine, while the decreased response may be due to vagal neuropathy.

Animals↗
PubMedDate not supplied

Differentiating small round cell sarcomas of the soft parts by an innovative immunogold labeling method: an ultrastructural study.

A new immunoelectron microscopy procedure was developed by remaking the fixed-frozen tissue specimens into LR White resin blocks suitable for postembedding colloidal gold immunolabeling, and used to examine 16 cases of small round cell soft tissue sarcomas. In rhabdomyosarcoma, ultrastructural double-immunogold staining demonstrated a coexpression of muscle specific actin and desmin in the same tumor cell. In both Ewing's sarcoma and peripheral neuroepithelioma, the heterogeneous expression of MIC2 gene product (p30/32MIC2) in each tumor cell was demonstrated as well. In peripheral neuroepithelioma, the colloidal gold immunolabeling for neurofilament demonstrated the intermediate filaments surrounding microtubules. The procedure for ultrastructural colloidal gold immunolabeling using fixed-frozen tissue is thus considered to be useful not only for tumor diagnosis, but also for investigating various subcellular structures.

Adolescent↗
PubMedDate not supplied

Angiogenesis does not correlate with prognosis or expression of vascular endothelial growth factor in synovial sarcomas.

To investigate the significance of tumour angiogenesis in synovial sarcoma on prognosis, we analysed the correlation between microvessel density (MVD) and various clinicopathological factors or immunohistochemical expression of vascular endothelial growth factor (VEGF) in 54 primary and recurrent synovial sarcomas. MVD in synovial sarcomas did not correlate with prognosis or VEGF expression. Furthermore, VEGF expression in synovial sarcomas did not have any prognostic value on overall survival. Our results indicate that angiogenesis does not play an important role in metastasis of synovial sarcoma and overall survival. Angiogenesis in synovial sarcoma may be controlled by angiogenesis activators other than VEGF.

Adolescent↗
PubMedDate not supplied

[Granulocyte colony-stimulating factor producing undifferentiated carcinoma of urinary bladder: a case report].

A case of bladder cancer producing granulocyte colony-stimulating factor (G-CSF) is reported. A 94-year-old woman with a progressive, grade 3 undifferentiated carcinoma, showed marked leukocytosis (maximum 29,780/mm3) with an elevated G-CSF (420 pg/ml). Immunohistochemical examination with monoclonal antibody specific for G-CSF revealed positive staining. Further examination for epidermal growth factor receptor (EGF-R) and p53 were both positive and Ki67 index were 40.7%. These data suggested that this tumor had extremely aggressive growing nature, as the biological character is this.

Aged↗
PubMedDate not supplied

Stimulation by nitric oxide of HCO3- secretion in bullfrog duodenum in vitro--roles of cyclooxygenase-1 and prostaglandins.

The effect of nitric oxide (NO) on HCO3- secretion was examined in vitro using the isolated preparation of bullfrog duodenum, in relation to cyclooxygenase (COX) isozymes and endogenous prostaglandins (PGs). The tissue was bathed in unbuffered Ringer solution gassed with 100% O2 on the mucosal side and HCO3- Ringer's solution gassed with 95% O2-5% CO2 on the serosal side. The HCO3- secretion was measured by a pH-stat method using 10 mM HCl as the titrant to keep the mucosal pH at 7.4. NOR-3 [(+/-)-(E)-Ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamine] was used as a NO donor and added to the serosal solution. To analyze the NOR-3 action, the effects of dibutyryl guanosine-3', 5'-cyclic monophosphate (dbcGMP), methylene blue, indomethacin (nonselective COX-inhibitor) and NS-398 (selective COX-2 inhibitor) on the HCO3- response were also examined. NOR-3 (1 x 10(-4) and 3 x 10(-4) M) caused an increase of HCO3- secretion in a dose-dependent manner, reaching the level of 2.5 times greater than basal values at 2 hr later. Likewise, dbcGMP (1 x 10(-3) M) also caused a significant increase of the duodenal HCO3- secretion. The HCO3- stimulatory action of NOR-3 was significantly attenuated by methylene blue (5 x 10(-5) M) and indomethacin (1 x 10(-5) M) but not by NS-398 (1 x 10(-5) M), and indomethacin also suppressed the HCO3- response to dbcGMP. The serosal release of PGE2 was significantly increased by both NOR-3 and dbcGMP, and these responses were inhibited by indomethacin but not NS-398. These results suggest that NO increases HCO3- secretion in Bullfrog duodenum in vitro, and this action is dependent on cGMP-related COX-1 activation and mediated by PGs.

16,16-Dimethylprostaglandin E2↗